Document Detail

Ameliorative effects of herbal combinations in hyperlipidemia.
Jump to Full Text
MedLine Citation:
PMID:  21941605     Owner:  NLM     Status:  MEDLINE    
The roots of Glycyrrhiza glabra, Withania somnifera, Asparagus racemosus, and Chlorophytum borivilianum and seeds of Sesamum indicum are ayurvedic medicinal plants used in India to treat several ailments. Our previous studies indicated that these plants possess hypolipidemic and antioxidant potential. The present study was aimed at investigating the composite effects of these plants on hypercholesterolemic rats. Three different combinations (5 gm%, given for four weeks) used in this study effectively reduced plasma and hepatic lipid profiles and increased fecal excretion of cholesterol, neutral sterol, and bile acid along with increasing the hepatic HMG-CoA reductase activity and bile acid content in hypercholesterolemic rats. Further, all three combinations also improved the hepatic antioxidant status (catalase, SOD, and ascorbic acid levels) and plasma total antioxidant capacity with reduced hepatic lipid peroxidation. Overall, combination I had the maximum effect on hypercholesterolemic rats followed by combinations II and III due to varying concentrations of the different classes of phytocomponents.
Nishant P Visavadiya; A V R L Narasimhacharya
Related Documents :
21696975 - The effect of ascorbic acid and dehydroascorbic acid on the root gravitropic response i...
23938205 - Ability of ellagic acid to alleviate osmotic stress on chickpea seedlings.
21941605 - Ameliorative effects of herbal combinations in hyperlipidemia.
6529855 - The plasma and erythrocyte fatty acid composition of poorly controlled, insulin-depende...
403285 - Beta-lactam antibiotics derived from nitrogen heterocyclic acetic acids. 1. penicillin ...
17622135 - Structural and coordination studies of "pearl oysterlike" porphyrins.
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2011-09-15
Journal Detail:
Title:  Oxidative medicine and cellular longevity     Volume:  2011     ISSN:  1942-0994     ISO Abbreviation:  Oxid Med Cell Longev     Publication Date:  2011  
Date Detail:
Created Date:  2011-09-23     Completed Date:  2012-01-17     Revised Date:  2013-06-27    
Medline Journal Info:
Nlm Unique ID:  101479826     Medline TA:  Oxid Med Cell Longev     Country:  United States    
Other Details:
Languages:  eng     Pagination:  160408     Citation Subset:  IM    
BRD School of Biosciences, Sardar Patel University, Vallabh Vidyanagar, India.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Ascorbic Acid / metabolism
Asparagus Plant / chemistry
Bile Acids and Salts / metabolism
Catalase / metabolism
Chlorophyta / chemistry
Feces / chemistry
Glycyrrhiza / chemistry
Hydroxymethylglutaryl CoA Reductases / metabolism
Hyperlipidemias / drug therapy*,  enzymology,  metabolism
Lipid Peroxidation / drug effects*
Lipids / blood
Liver / drug effects,  enzymology,  metabolism
Plant Extracts / chemistry,  pharmacology*,  therapeutic use*
Sesamum / chemistry
Superoxide Dismutase / metabolism
Withania / chemistry
Reg. No./Substance:
0/Bile Acids and Salts; 0/Lipids; 0/Plant Extracts; 50-81-7/Ascorbic Acid; EC 1.1.1.-/Hydroxymethylglutaryl CoA Reductases; EC; EC Dismutase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Full Text
Journal Information
Journal ID (nlm-ta): Oxid Med Cell Longev
Journal ID (publisher-id): OXIMED
ISSN: 1942-0900
ISSN: 1942-0994
Publisher: Hindawi Publishing Corporation
Article Information
Download PDF
Copyright © 2011 N. P. Visavadiya and A. V. R. L. Narasimhacharya.
Received Day: 12 Month: 5 Year: 2011
Accepted Day: 15 Month: 7 Year: 2011
Print publication date: Year: 2011
Electronic publication date: Day: 15 Month: 9 Year: 2011
Volume: 2011E-location ID: 160408
ID: 3173889
PubMed Id: 21941605
DOI: 10.1155/2011/160408

Ameliorative Effects of Herbal Combinations in Hyperlipidemia
Nishant P. Visavadiya1, 2I2
A. V. R. L. Narasimhacharya1*
1BRD School of Biosciences, Sardar Patel University, Sardar Patel Maidan, Vadtal Road, Satellite Campus, P.O. Box 39, Vallabh Vidyanagar 388 120, India
2Spinal Cord and Brain Injury Research Center (SCoBIRC), University of Kentucky, Biomedical & Biological Sciences Research Building B0436-41, 741 S. Limestone St., Lexington, KY 40536, USA
Correspondence: *A. V. R. L. Narasimhacharya:
[other] Academic Editor: Gabriele Saretzki

1. Introduction

Hypercholesterolemia is one of the major risk factors that precipitate coronary heart disease (CHD) and atherosclerosis [1]. Besides medication, composition of the diet also plays an important role in the management of lipid and lipoprotein concentrations in blood. Plant-based therapies are recognized for their therapeutic applications as they either have minimal or no side effects [1, 2]. Over the past 20 years or so, interest in traditional medicines has increased considerably in many parts of the world. Traditional medicines all over the world are being reevaluated by extensive research on different plant species with regard to their therapeutic principles and potential. Despite the progress in conventional chemistry and pharmacology in producing effective drugs, the plant kingdom might yet provide useful sources of new medicines. Plants produce an amazing variety of metabolites such as isoflavones, phytosterols, saponins, fibers, polyphenols, flavonoids, and ascorbic, and these have aroused much interest for their role in lipid and antioxidant metabolism [37].

Several herbal combinations have been tested experimentally for their potential in ameliorating various ailments, for example, diabetes, allergic rhinitis, atherosclerosis, rheumatoid arthritis, and antimicrobial activity [712]. For instance, an experimental combination (of roots of ashwagandha, rhizomes of ginger, and young mulberry leaves) treatment given to NIDDM human subjects revealed a significant reduction in blood glucose, total cholesterol, triglycerides, LDL cholesterol and VLDL cholesterol [13].

A number of polyherbal formulations to treat various disorders are also being marketed by several reputed ayurvedic drug manufacturers in India which include among others Chyawanprash, Active Blood Purifier (44 and 7 herbs, resp.; Dabur India Ltd.), D-400, Liv 52, OST-6, Cystone (32, 7, 5, and 7 herbs, resp.; Himalaya Drug Co.), Livergen (6 herbs; Standard Pharmaceuticals), Livokin (17 herbs; Herbo-Med), Stimuliv (4 herbs; Franco-Indian Pharmaceuticals Pvt. Ltd.), Tefroliv (9 herbs; TTK Pharma Pvt. Ltd.), Pankajakasthuri (14 herbs; Herbals India Pvt. Ltd.), and Dihar (8 herbs; Rajsha Pharmaceuticals).

The present investigation is aimed at finding an effective combination of different proportions of certain medicinal plants which could supplement as a therapeutic agent for hyperlipidaemia. The plants used in the present investigation, that is, Glycyrrhiza glabra (F. Fabaceae), Withania somnifera (F. Solanaceae), Asparagus racemosus (F. Liliaceae), Chlorophytum borivilianum (F. Liliaceae), and Sesamum indicum (F. Pedaliaceae), have ethnobotanical history and are widely used in Ayurvedic System of Medicine in India for various ailments. Our previous work on G. glabra, C. borivilianum, W. somnifera, S. indicum, and A. racemosus showed dose-dependent (5 and 10 gm% of roots/seed powder) beneficial effects on hypercholesterolemic rats [1418]. We found that G. glabra was the most potent of the plants studied with reference to hypolipidaemic/hypocholesterolemic and antioxidant effects followed by W. somnifera, A. racemosus, C. borivilianum, and S. indicum. In this study, we have investigated the effect of different combinations of these plants on lipid profiles, fecal-cholesterol, neutral sterol, and bile acid excretion patterns and the consequent effects on hepatic cholesterol and bile acid production in hypercholesterolemic rats. An attempt was also made to study the status of hepatic lipid peroxidation, the profiles of antioxidants-catalase, superoxide dismutase, and ascorbic acid as well as ferric reducing ability of the plasma (FRAP) in hypercholesterolemic rats.

2. Results
2.1. Phytoconstituents and FRAP Value in Different Combinations

The quantitative phytochemical analysis of all three combinations (C-I, C-II, and C-III) indicated that these combinations contained fiber, phytosterols, saponin, polyphenol, flavonoids, and ascorbic acid. The FRAP values as total antioxidant concentrations in combinations I, II, and III indicated the highest value for C-I followed by C-II and C-III (Table 2).

2.2. Body Weight, Food Intake, and Liver Weight

There were no significant differences in food intake or body weights in any group (NC, HC, C-I, C-II, and C-III). However, liver weight decreased significantly (P < 0.002) in C-I (15%), C-II (14%), and C-III (12%) groups compared to HC animals (data not presented).

2.3. Plasma and Hepatic Lipid Profiles

The plasma lipid profiles significantly decreased (P < 0.002) with all three combinations. All three groups (C-I, C-II, and C-III) registered significant decline in plasma TL (C-I: 30%; C-II: 32%; C-III: 20%), TC (40%; 43%; 29%), TG (28%; 15%; 14%), and LDL (53%; 57%; 37%) levels, and an increase in HDL-C (35%; 37%; 21%, P < 0.002) was noted in these groups as compared to HC group (Figure 1). Very-low-density lipoprotein (VLDL) levels in the test groups (C-I: 7.8 ± 0.2; C-II: 9.2 ± 0.2; C-III: 9.3 ± 0.2 mg/dL, P < 0.002) were significantly lower than in HC group (10.8 ± 0.2 mg/dL). Thus, the combination diets reduced the VLDL concentrations by 28%, 15%, and 14%, respectively. Similarly, the atherogenic index (AI) was also reduced in the combination-diet-treated groups (C-I: 3.3 ± 0.1; C-II: 3.1 ± 0.1; C-III: 4.4 ± 0.1, P < 0.002) as compared to HC group (7.6 ± 0.3). The hepatic lipid profiles too significantly declined (P < 0.002) in the test groups: TL (by 28%, 34%, and 20% in C-I, C-II, and C-III, resp.) and TC (by 31%, 35%, and 20% in C-I, C-II, and C-III, resp.) but not significantly TG (by 17%, 9%, and 8% in C-I, C-II, and C-III resp., P < 0.002) as compared to HC group (Table 3).

2.4. Hepatic HMG-CoA Reductase and Bile Acids Levels

Administration of different combinations to hypercholesterolemic rats resulted in a significant increase in (P < 0.002) hepatic HMG-CoA reductase activity (C-I, C-II, and C-III groups: 16%, 19%, and 13%, resp.) as compared to HC group. Hepatic bile acid content also increased (P < 0.002, 27%; 33%; 16%) in these groups as compared to those of hypercholesterolemic control group (Table 3).

2.5. Fecal Cholesterol, Neutral Sterols, and Bile Acid Content

There were increments (P < 0.002) in fecal cholesterol (15%; 19%; 11%), neutral sterol (19%; 23%; 12%) and bile acid (29%; 34%; 18%) excretion in C-I, C-II, and C-III groups as compared to HC group (Table 4).

2.6. Hepatic Lipid Peroxidation and Antioxidant Profiles

The hepatic lipid peroxidation decreased significantly (P < 0.002) in all three groups (C-I: 33%, C-II: 26%, and C-III: 17%) when compared to that of HC group. Both catalase (P < 0.002, C-I: 36%, C-II: 32%, and C-III: 14%) and SOD (P < 0.002, C-I: 27%, C-II: 20%, and C-III: 19%) activities also increased in these groups significantly compared to those in HC group (Figure 2). The C-I, C-II, and C-III group rats registered had increased hepatic ascorbic acid content (132.7 ± 3.8; 114.3 ± 3.1; 110.3 ± 3.2 μg/g, P < 0.002) as compared to the hypercholesterolemic (95.5 ± 2.8 μg/g) rats. FRAP values increased in all groups that is, C-I, CII, and C-III (P < 0.002, 35%; 29%; 21%, resp.), as compared to HC group (Figure 1).

3. Discussion

The present investigation demonstrates that the plant combinations used in this study are effective in lowering cholesterol levels in hypercholesterolemic rats. Plant metabolites, the sterols, soybean proteins, and fibers are reported to reduce the risk of CHD through their cholesterol lowering properties, independently and also in combination [8]. In this study, the observed cholesterol-lowering effects of different plant combinations administered to hypercholesterolemic rats could be related to an increased excretion of cholesterol, neutral sterols, and bile acid. The phytochemical analysis of the combinations (C-I, C-II, and C-III) revealed the presence of fibers, saponins, phytosterols, polyphenols, flavonoids, and ascorbic acid among which phytosterols, fibers as well as saponins could be important in cholesterol elimination in hypercholesterolemic rats administered with different combinations. These components have received considerable attention for their plasma cholesterol-lowering effects [3, 1921]. It is known that phytosterols have a greater affinity for micelles than cholesterol because of their greater hydrophobicity. Therefore, they can easily displace intestinal cholesterol from the micelles, reducing intestinal cholesterol absorption, and consequently reduce hepatic and plasma cholesterol concentrations [21, 22]. Dietary fibers appear to interfere with cholesterol absorption and its enterohepatic bile circulation resulting in depletion of hepatic cholesterol pools and alteration in lipoprotein metabolism. Besides, the cholesterol lowering effect of fibers is considered primarily due to an increased excretion of cholesterol and bile acids [19, 23]. Saponins are also capable of precipitating cholesterol from micelles and interfering with enterohepatic circulation of bile acids making it unavailable for intestinal absorption and hence reduce plasma cholesterol levels [20, 24]. It is noteworthy that the excretion of cholesterol, neutral sterol, and bile acid was significantly higher with administration of different plant combinations when compared to that of hypercholesterolemic rats. These losses of fecal cholesterol and bile acid could have depleted hepatic cholesterol levels, and, as a consequence, a compensatory increase in hepatic cholesterogenesis and bile acid synthesis was noted with demonstrably higher hepatic HMG-CoA reductase activity and bile acid production in different combination groups as compared to those of hypercholesterolemic control rats. A significant decline in plasma LDL-cholesterol along with increased bile acid production in different combination groups could also be due to the fiber and saponin content of combinations as both fibers and saponins are reported to lower plasma LDL-cholesterol levels via interruption of cholesterol and bile acid absorption and increased LDL-receptor activity; the LDL-cholesterol could then be subsequently catabolized to bile acid [23, 25].

Rats fed different combinations had lower hepatic and plasma TG concentrations than rats fed cholesterol alone. The TG depletion may be related to the ability of fiber to inhibit absorption of lipids from the intestinal lumen [23]. Moreover, saponins are reported to inhibit pancreatic lipase activity and reduce the TG concentrations [26] leading to subsequent reduction in the VLDL-C concentration in plasma [22]. Presently all three experimental groups (C-I, C-II and C-III) showed a simultaneous decline in plasma TG and VLDL-C levels that could be correlated with the fiber and saponin content of the experimental diets. Several studies documented that low levels of HDL-cholesterol are indicative of high risk for CHD and an increase in HDL-C level is considered beneficial [1]. Epidemiological studies have also shown that high HDL-cholesterol levels could potentially contribute to antiatherogenecity, inhibit LDL-oxidation and protect endothelial cells from the cytotoxic effects of oxidized LDL [27]. Presently noted high levels of plasma HDL-cholesterol in hypercholesterolemic animals administered with different combinations as compared to HC animals indicate the physiological role played by these combinations in elevating HDL-cholesterol levels. While dietary saponins and fibers are not known to elevate HDL-cholesterol levels [20, 23], ascorbate and flavonoids are reported to increase the HDL-cholesterol concentrations [28]. As the combinations contained both ascorbic acid and flavonoids, the increased HDL-cholesterol levels in hypercholesterolemic animals administered with combination diets could be attributed to these plant metabolites.

Recent studies have demonstrated a direct relationship between hypercholesterolemia and CHD [1]. The dietary cholesterol during its metabolism is delivered to the hepatic cells where substantial amounts of reactive oxygen species are generated [29]. This process is believed to generate highly toxic products, including lipid peroxides as aldehydes, epoxides and carbonyls, and cause rapid consumption of antioxidants such as vitamin E or vitamin C [29]. Further, high cholesterol diet increases serum LDL levels, and, due to oxidative stress, the LDL is oxidized increasingly thereby facilitating atherosclerotic plaque formation [30]. Many studies have shown that dietary polyphenols, flavonoids, carotenoids, vitamin C, and lignans can effectively scavenge free radicals, reduce the levels of lipid peroxidation and oxidation of LDL and, prevent the progress of atherosclerosis [3, 28, 3032]. The polyphenols with their antioxidant activity, chiefly due to their redox properties, act as reducing agents, hydrogen donors, and singlet oxygen quenchers with a metal chelating potential [3, 32]. The polyphenols and flavonoids are reported to scavenge free radicals, including hydroxyl and superoxide anions, stimulate SOD and catalase gene expression, reduce the malondialdehyde concentrations, and improve lipid profiles in experimental animals [10, 32, 33]. Ascorbic acid is a major chain-breaking antioxidant against free radicals and conjugates with cytotoxic, genotoxic, and lipid peroxidation products to eliminate them [34]. The root extract of G. glabra has antioxidant activity reportedly due to the presence of a variety of phenolic compounds including flavonoids, isoflavonoids, chalcones, and bibenzyls [35]. The antioxidant and antiperoxidative properties of sesame seed are related to lignans, such as sesamol, sesamolinol, pinoresinol, and sesaminol [31]. The sitoindosides VII-X and Withaferin A (Glycowithanolides) of W. somnifera have been implicated in increasing the antioxidant activity in rat brain and stimulated catalase and SOD activities [36]. In the present study, high cholesterol (HC) fed rats showed a remarkable decline in antioxidant status and concurrently increased lipid peroxidation as compared to the normal control (NC) rats. However, all three combinations treated hypercholesterolemic rats indicate considerable decrease in lipid peroxidation and improvement in the antioxidant status. Further, the aqueous extract and plasma FRAP-values as well as hepatic catalase, SOD and ascorbic acid levels of C-I were highest compared to C-II and C-III groups. The malondialdehyde level in C-I group also showed a significant decline as compared to C-II and C-III groups. The phytochemical analyses of combinations have indicated the presence of polyphenols, flavonoids, and ascorbic acid. Consequently, these elevated levels of hepatic antioxidants in combinations-treated rats might be due to the presence of polyphenols and flavonoids in the experimental diet. In this context, our recent in vitro studies on aqueous and ethanolic extracts of G. glabra, A. racemosus, C. borivilianum, and S. indicum demonstrated that these plants possess considerable free radical scavenging potency against superoxide, nitric oxide, and hydroxyl radicals due to phenolic content of plants [3740]. In addition, these plants markedly suppressed the metal-catalyzed lipid peroxidation in hepatic mitochondrial fractions isolated from rats as well as favorably affected atherosclerosis risk status by reducing human serum and LDL oxidation caused by copper ion [3740]. Taken together, presently observed high antioxidant status in combinations-treated hypercholesterolemic rats may be due to free radical scavenging action exerted by combinations diet. Additionally, the combinations could be a potent source for ascorbic acid as it suppressed the propagation of lipid peroxidation and reduced the levels of malondialdehyde. Unlike our earlier reports on these individual plants [1418], the combinations used in the present investigation (C-I, C-II, and C-III) are with much lower levels of additions of individual plants (ranging between 0.625 and 1.5 gm as against 5 and 10 gm%) in the diet (Table 1). Yet these combinations significantly reduced the lipid profiles and improved the body antioxidant capacity of hypercholesterolemic rats. Thus, the herbal combinations-C-I, C-II, and C-III, exhibited differential hypocholesterolemic and antioxidant activities when fed to hypercholesterolemic rats. This could be due to varying concentrations of different classes of phytocomponents in the combinations such as fiber, phytosterols, saponins, polyphenols, flavonoids and vitamin C (Table 2). In conclusion, although all the three tested combinations of plants were effective as hypocholesterolemic and antioxidant agents, C-I appeared to be more potent in reducing body lipid profiles and lipid peroxidation and improving the antioxidant status of hypercholesterolemic rats.

4. Materials and Methods
4.1. Preparation of Plant Combinations and Phytochemical Analysis

The roots of Glycyrrhiza glabra (GG) and seeds of Sesamum indicum (SI) were purchased from the local merchandise and roots of Withania somnifera (WS), Asparagus racemosus (AR) and Chlorophytum borivilianum (CB) were collected from University Botanical Gardens and authenticated by our faculty Taxonomist Dr. A. S. Reddy. Roots and seeds were air-dried completely, ground to powder, and used for preparation of different combinations (i.e., combinations I, II, and III). All combinations were prepared using a 5 gm% mixture of roots/seed powder of the plants (Table 1).

Phytoconstituents of C-I, C-II, and C-III were quantified by standard methods. The total fiber, polyphenol, and flavonoid were estimated according to Thimmaiah [41]. Phytosterol and saponin contents of the combinations were estimated using ferric chloride-sulphuric acid and vanillin-sulphuric acid methods, respectively, with β-sitosterol and saponin as standards [42, 43]. The total ascorbic acid content was estimated using 2,4-dinitrophenyl hydrazine reagent [44]. The ferric-reducing ability of combinations (FRAP) were measured as the concentrations of total antioxidants using the TPTZ (2,4,6-tripyridyl-s-triazine) HCl-FeCl3 reagent at 593 nm. Calibration curve of ferrous sulfate was used [45].

4.2. Animals

Male albino rats (Charles Foster, bred in the Department's Animals House, weighing 150–200 gm) were housed individually with ad libitum access to water and fed on commercial feed (Pranav Agro Ind. Ltd., Pune, India) in a well-ventilated animal unit (26 ± 2°C, humidity 56%, 12 hour-light/dark cycle). The care and procedures adopted for the present investigation were in accordance with rules and regulations of CPCSEA, and the experiment was approved by Institutional Animal Ethics Committee (IAEC).

4.3. Experimental Design

After a 10-day adaptation period, 30 rats were divided into 5 groups of 6 rats each: NC: normal control rats receiving only commercial feed/ basal diet, HC: high cholesterol diet control rats receiving basal diet with 0.5 gm% cholesterol and 1.0 gm% sodium taurocholate, C-I: rats receiving high cholesterol diet + combination I, C-II: rats receiving high cholesterol diet + combination II, and C-III: rats receiving high cholesterol diet + combination III. The basal diet contained 55.67 gm% carbohydrates, 22.12 gm% protein, 4.06 gm% fat, 3.76 gm% fiber, 5.64 gm% mineral mixture, and 8.75% moisture content.

At the end of the four-week treatment, 24-hour fecal samples were collected from individual cages. Animals were fasted overnight and sacrificed under mild anesthesia (diethyl ether). Blood was collected by cardiac puncture, and plasma was separated by centrifugation. Liver was excised, and, both plasma and liver were kept frozen until analyzed.

4.4. Plasma and Hepatic Lipid Profiles

Plasma total lipid (TL) content was estimated by sulpho-phosphovanillin method [46]. Plasma cholesterol (TC), HDL-cholesterol (HDL-C), and triglycerides (TG) were assayed using commercial diagnostic kits (Eve's Inn Diagnostics, Vadodara, India). Low-density lipoprotein cholesterol (LDL-C), very-low-density lipoprotein cholesterol (VLDL-C), and atherogenic index (AI) were calculated according to Friedewald equations [47]. The liver TL was extracted in chloroform : methanol (2 : 1) [48] and estimated gravimetrically. Hepatic TC and TG were extracted [48] and estimated using the standard kits (Eve's Inn Diagnostics, Vadodara, India).

4.5. Hepatic HMG-CoA Reductase and Bile Acid Profile

Hepatic HMG-CoA reductase (EC activity was measured in terms of the ratio of HMG-CoA to mevalonate using hydroxylamine-FeCl3 reagent [49]. Briefly, the hepatic HMG-CoA was determined by its reaction with hydroxylamine reagent at alkaline pH and followed by colorimetric measurement of the resulting hydroxamic acid by formation of complexes with ferric salts at 540 nm. The hepatic mevalonate was estimated by reaction with the same reagent but at pH 2.1. At this pH, the lactone form of mevalonate readily reacts with hydroxylamine to form the hydroxamate. Thus, hepatic HMG-CoA and mevalonate concentration in the tissue homogenate are estimated in terms of absorbances, and the ratio between the two is taken as an index of the HMG-CoA reductase enzyme activity that catalyzes the conversion of 3-hydroxy-3-methylglutaryl-coenzyme A to mevalonate. The ratio is inversely proportional to the enzyme activity, that is, the increase in ratio corresponds to a decrease in enzyme activity. The alkaline-ethanol extract of hepatic bile acid was acidified and estimated using vanillin-phosphoric acid reagent [50].

4.6. Fecal Cholesterol, Neutral Sterol, and Bile Acid Content

The fecal cholesterol, neutral sterol, and bile acid were extracted using alkaline methanol medium [51], and fecal cholesterol and neutral sterol contents were estimated [52] using standard kits (Eve's Inn Diagnostics, Vadodara, India). A portion of the extract was acidified and used for bile acid estimation [50].

4.7. Hepatic Lipid Peroxidation and Antioxidant Profile

The concentration of the lipid peroxide products of hepatic TBARS (Thiobarbituric acid reactive substances) was measured by TBA-TCA-HCl reagent (0.37% TBA, 15% TCA, 0.25 N HCl) at 532 nm using a molar absorption coefficient of 1.56 × 105 M−1cm−1 [53]. Catalase (EC activity was measured spectrophotometrically as decomposition of H2O2 at 240 nm [54]. The superoxide dismutase (SOD; EC was assayed by using NADPH and PMS (phenazine methosulfate) reagents under nonacidic conditions, which reduces NBT (nitroblue tetrazolium salt) and forms a blue coloured formazon that can be measured at 560 nm [55]. Total ascorbic acid was determined using 2,4-dinitro phenyl hydrazine (DNPH) H2SO4 reagent [44]. The FRAP value of plasma was measured by the method of Benzie and Strain [45].

4.8. Statistical Evaluation

Results are expressed as means ± SEM. Significant differences among the groups were determined by one-way ANOVA using the 10th version of SPSS with Duncan's test as post hoc analysis. Differences were considered significant if P < 0.002.

Conflict of Interests

The authors declare that they have no conflict of interests.


The financial assistance in the form of a research grant (A. V. R. L. Narasimhacharya) and project fellow (N. P. Visavadiya) from U.G.C., New Delhi, India is gratefully acknowledged. The authors are especially thankful to Miss Rupal A. Vasant, Research Fellow, for her help in paper preparation.

1. Mensink RP,Aro A,Hond ED,et al. PASSCLAIM—diet-related cardiovascular diseaseEuropean Journal of NutritionYear: 2003421627
2. Singh B,Bhat TK,Singh B. Potential therapeutic applications of some antinutritional plant secondary metabolitesJournal of Agricultural and Food ChemistryYear: 200351195579559712952405
3. Rice-Evans CA,Miller NJ,Bolwell PG,Bramley PM,Pridham JB. The relative antioxidant activities of plant-derived polyphenolic flavonoidsFree Radical ResearchYear: 19952243753837633567
4. Kerckhoffs DAJM,Brouns F,Hornstra G,Mensink RP. Effects on the human serum lipoprotein profile of β-glucan, soy protein and isoflavones, plant sterols and stanols, garlic and tocotrienolsJournal of NutritionYear: 200213292494250512221200
5. Tripathi UN,Chandra D. The plant extracts of Momordica charantia and Trigonella foenum graecum have antioxidant and anti-hyperglycemic properties for cardiac tissue during diabetes mellitusOxidative Medicine and Cellular LongevityYear: 20092529029620716916
6. Verma S,Gupta ML,Dutta A,Sankhwar S,Shukla SK,Flora SJS. Modulation of ionizing radiation induced oxidative imbalance by semi-fractionated extract of Piper betle: an in vitro and in vivo assessmentOxidative Medicine and Cellular LongevityYear: 201031445220716927
7. Bulku E,Zinkovsky D,Patel P,et al. A novel dietary supplement containing multiple phytochemicals and vitamins elevates hepatorenal and cardiac antioxidant enzymes in the absence of significant serum chemistry and genomic changesOxidative Medicine and Cellular LongevityYear: 20103212914420716937
8. Lamarche B,Desroches S,Jenkins DJA,et al. Combined effects of a dietary portfolio of plant sterols, vegetable protein, viscous fibre and almonds on LDL particle sizeBritish Journal of NutritionYear: 200492465766315522135
9. Said O,Fulder S,Khalil K,Azaizeh H,Kassis E,Saad B. Maintaining a physiological blood glucose level with “glucolevel”, a combination of four anti-diabetes plants used in the traditional Arab herbal medicineEvidence-Based Complementary and Alternative MedicineYear: 20085442142818955212
10. Mothana RAA,Abdo SAA,Hasson S,Althawab FMN,Alaghbari SAZ,Lindequist U. Antimicrobial, antioxidant and cytotoxic activities and phytochemical screening of some yemeni medicinal plantsEvidence-Based Complementary and Alternative MedicineYear: 20107332333018955315
11. Tripathi YB,Singh BK,Pandey RS,Kumar M. BHUX: a patent herbal formulation to prevent atherosclerosisEvidence-Based Complementary and Alternative MedicineYear: 20052221722115937563
12. Nozaki K,Hikiami H,Goto H,Nakagawa T,Shibahara N,Shimada Y. Keishibukuryogan (Gui-Zhi-Fu-Ling-Wan), a Kampo formula, decreases disease activity and soluble vascular adhesion molecule-1 in patients with rheumatoid arthritisEvidence-Based Complementary and Alternative MedicineYear: 20063335936416951720
13. Andallu B,Radhika B,Suryakantham V. Effect of aswagandha, ginger and mulberry on hyperglycemia and hyperlipidemiaPlant Foods for Human NutritionYear: 20035831712859008
14. Visavadiya NP,Narasimhacharya AVRL. Hypocholesterolaemic and antioxidant effects of Glycyrrhiza glabra (Linn) in ratsMolecular Nutrition and Food ResearchYear: 200650111080108617054099
15. Visavadiya NP,Narasimhacharya AVRL. Ameliorative effect of Chlorophytum borivilianum root on lipid metabolism in hyperlipaemic ratsClinical and Experimental Pharmacology and PhysiologyYear: 200734324424917250646
16. Visavadiya NP,Narasimhacharya AVRL. Hypocholesteremic and antioxidant effects of Withania somnifera (Dunal) in hypercholesteremic ratsPhytomedicineYear: 2007142-313614216713218
17. Visavadiya NP,Narasimhacharya AVRL. Sesame as a hypocholesteraemic and antioxidant dietary componentFood and Chemical ToxicologyYear: 20084661889189518353516
18. Visavadiya NP,Narasimhacharya AVRL. Asparagus root regulates cholesterol metabolism and improves antioxidant status in hypercholesteremic ratsEvidence-Based Complementary and Alternative MedicineYear: 20096221922618955232
19. Moundras C,Behr SR,Rémésy C,Demigné C. Fecal losses of sterols and bile acids induced by feeding rats guar gum are due to greater pool size and liver bile acid secretionJournal of NutritionYear: 19971276106810769187619
20. Harwood HJ Jr.,Chandler CE,Pellarin LD,et al. Pharmacologic consequences of cholesterol absorption inhibition: alteration in cholesterol metabolism and reduction in plasma cholesterol concentration induced by the synthetic saponin β-tigogenin cellobioside (CP- 88818; tiqueside)Journal of Lipid ResearchYear: 19933433773958468523
21. Ostlund RE Jr.. Phytosterols and cholesterol metabolismCurrent Opinion in LipidologyYear: 2004151374115166807
22. Howell TJ,MacDougall DE,Jones PJH. Phytosterols partially explain differences in cholesterol metabolism caused by corn or olive oil feedingJournal of Lipid ResearchYear: 19983948929009555952
23. Venkatesan N,Devaraj SN,Devaraj H. Increased binding of LDL and VLDL to apo B,E receptors of hepatic plasma membrane of rats treated with FibernatEuropean Journal of NutritionYear: 200342526227114569407
24. Francis G,Kerem Z,Makkar HPS,Becker K. The biological action of saponins in animal systems: a reviewBritish Journal of NutritionYear: 200288658760512493081
25. Rajendran S,Deepalakshmi PD,Parasakthy K,Devaraj H,Devaraj SN. Effect of tincture of Crataegus on the LDL-receptor activity of hepatic plasma membrane of rats fed an atherogenic dietAtherosclerosisYear: 19961231-22352418782854
26. Han LK,Zheng YN,Xu BJ,Okuda H,Kimura Y. Saponins from platycodi radix ameliorate high fat diet-induced obesity in miceJournal of NutritionYear: 200213282241224512163669
27. Assmann G,Nofer JR. Atheroprotective effects of high-density lipoproteinsAnnual Review of MedicineYear: 200354321341
28. Vinson JA,Hu SJ,Jung S,Stanski AM. A citrus extract plus ascorbic acid decreases lipids, lipid peroxides, lipoprotein oxidative susceptibility, and atherosclerosis in hypercholesterolemic hamstersJournal of Agricultural and Food ChemistryYear: 199846414531459
29. Erdinçler DS,Seven A,Inci F,Beğer T,Candan G. Lipid peroxidation and antioxidant status in experimental animals: effects of aging and hypercholesterolemic dietClinica Chimica ActaYear: 199726517784
30. Warnholtz A,Mollnau H,Oelze M,Wendt M,Münzel T. Antioxidants and endothelial dysfunction in hyperlipidemiaCurrent Hypertension ReportsYear: 200131536011177709
31. Kang MH,Naito M,Sakai K,Uchida K,Osawa T. Mode of action of sesame lignans in protecting low-density lipoprotein against oxidative damage in vitroLife SciencesYear: 199966216117110666012
32. Pandey KB,Rizvi SI. Plant polyphenols as dietary antioxidants in human health and diseaseOxidative Medicine and Cellular LongevityYear: 20092527027820716914
33. Ranaivo HR,Rakotoarison O,Tesse A,et al. Cedrelopsis grevei induced hypotension and improved endothelial vasodilatation through an increase of Cu/Zn SOD protein expressionAmerican Journal of PhysiologyYear: 20042862H775H78114551050
34. Sowell J,Frei B,Stevens JF. Vitamin C conjugates of genotoxic lipid peroxidation products: structural characterization and detection in human plasmaProceedings of the National Academy of Sciences of the United States of AmericaYear: 200410152179641796915608056
35. Fuhrman B,Buch S,Vaya J,et al. Licorice extract and its major polyphenol glabridin protect low-density lipoprotein against lipid peroxidation: in vitro and ex vivo studies in humans and in atherosclerotic apolipoprotein E-deficient miceAmerican Journal of Clinical NutritionYear: 19976622672759250104
36. Bhattacharya SK,Satyan KS,Ghosal S. Antioxidant activity of glycowithanolides from Withania somniferaIndian Journal of Experimental BiologyYear: 19973532362399332168
37. Visavadiya NP,Soni B,Dalwadi N. Evaluation of antioxidant and anti-atherogenic properties of Glycyrrhiza glabra root using in vitro modelsInternational Journal of Food Sciences and NutritionYear: 200960213514919384750
38. Visavadiya NP,Soni B,Soni B,Madamwar D. Suppression of reactive oxygen species and nitric oxide by Asparagus racemosus root extract using in vitro studiesCellular and Molecular BiologyYear: 200955OL1083OL109519267991
39. Visavadiya NP,Soni B,Dalwadi N,Madamwar D. Chlorophytum borivilianum as potential terminator of free radicals in various in vitro oxidation systemsDrug and Chemical ToxicologyYear: 201033217318220307144
40. Visavadiya NP,Soni B,Dalwadi N. Free radical scavenging and antiatherogenic activities of Sesamum indicum seed extracts in chemical and biological model systemsFood and Chemical ToxicologyYear: 200947102507251519607871
41. Thimmaiah SK. Standard Methods of Biochemical AnalysisYear: 1999New Delhi, IndiaKalyani
42. Goad LJ,Akihisa T. Analysis of SterolsYear: 19971st editionLondon, UKBlackie Academic and Professional, an imprint of Chapman and Hall
43. Ebrahimzadeh H,Niknam V. A revised spectrophotometric method for determination of triterpenoid saponinsIndian DrugsYear: 1998356379381
44. Schaffert RR,Kingsley GR. A rapid, simple method for the determination of reduced, dehydro-, and total ascorbic acid in biological materialThe Journal of Biological ChemistryYear: 19552121596813233208
45. Benzie IFF,Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assayAnalytical BiochemistryYear: 1996239170768660627
46. Frings CS,Fendley TW,Dunn RT,Queen CA. Improved determination of total serum lipids by the sulfo-phospho-vanillin reactionClinical ChemistryYear: 19721876736745037917
47. Friedewald WT,Levy RI,Fredrickson DS. Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifugeClinical ChemistryYear: 19721864995024337382
48. Folch J,Lees M,Sloane Stanley GH. A simple method for the isolation and purification of total lipides from animal tissuesThe Journal of Biological ChemistryYear: 1957226149750913428781
49. Rao AV,Ramakrishnan S. Indirect assessment of hydroxymethylglutaryl CoA reductase (NADPH) activity in liver tissueClinical ChemistryYear: 19752110152315251157326
50. Snell FD,Snell CT. Colorimetric Methods of AnalysisYear: 19533New York, NY, USAD. Van Nostrand
51. Kalek HD,Stellaard F,Kruis W,Paumgartner G. Detection of increased bile acid excretion by determination of bile acid content in single stool samplesClinica Chimica ActaYear: 198414018590
52. Snell FD,Snell CT. Colorimetric Methods of AnalysisYear: 19544New York, NY, USAD. Van Nostrand
53. Buege JA,Aust SD. [30] Microsomal lipid peroxidationMethods in EnzymologyYear: 197852302310672633
54. Aebi H. Bergmeyer HUCatalaseMethods of Enzymatic AnalysisYear: 197422nd editionNew York, NY, USAAcademic Press673684
55. Kakkar P,Das B,Viswanathan PN. A modified spectrophotometric assay of superoxide dismutaseIndian Journal of Biochemistry and BiophysicsYear: 1984212130132

Article Categories:
  • Research Article

Previous Document:  Targeting HDACs: a promising therapy for Alzheimer's disease.
Next Document:  Evaluation of chromosomal instability in diabetic rats treated with naringin.